论文部分内容阅读
荧光原位杂交(fluorescence in situ hybridization,FISH)是一种使用非放射性探针直接在染色体、细胞或组织水平上检测和定位靶核苷酸序列的分子细胞遗传学技术.本研究以紫花苜蓿的重复序列E180、MsCR-3和MsTR-1为模板设计了Oligo-MsCR3、Oligo-MsTR1和Oligo-E 180三个新寡核苷酸序列;并使用荧光原位杂交方法对这三个寡核苷酸序列在紫花苜蓿染色体上的分布进行了定位.结果表明:本研究所开发的3种新的寡核苷酸探针Oligo-MsCR3、Oligo-MsTR1和Oligo-E 180可以直接用于紫花苜蓿的荧光原位杂交分析.并且,结合染色体相对长度及臂比值信息,这3个探针可以对紫花苜蓿的16对染色体进行有效识别.因此,本研究为世界性优良牧草“紫花苜蓿”建立了一个更加简单、方便的细胞遗传学分析方法.“,”Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that non-radioactive probes are used to directly detect and localize target nucleotide sequences on chromosomes,cells,or tissue.In this study,three new oligonucleotide probes,Oligo-MsCR3,Oligo-MsTR1 and Oligo-E180 were designed based on the repetitive sequences E180,MsCR-3 and MsTR-1 of alfalfa.The distributions of Oligo-MsCR3,Oligo-MsTR1 and Oligo-E 180 on alfalfa chromosomes were detected by FISH.The results showed that these three oligonucleotide probes can be used for FISH analysis.Combined with relative lengths and arm ratios of chromosomes,these three probes could effectively identify 16 pairs of chromosomes of alfalfa.Therefore,our study has established a simpler and more convenient cytogenetic method for the world's most important forage legume “Alfalfa”.